This Program Project is the continuation of collaboration between three research groups that have been working together since 1993. The successful interdisciplinary team represents knowledge in chemical collaborative group has focused on the development of microfabricated DNA analysis devices with complete process integration. The continuation proposal will develop a fully integrated DNA sample handling and analysis technology for genotyping and sequencing. All the steps for extracting genetic information will occur within a single self-contained system having minimal operator interaction. The microfluidic, optical, and electronic components of the system are fabricated on silicon, silicon/glass, or silicon/polymer substrates, yielding a micromechanical integrated DNA analysis technology. To ensure inexpensive design, testing, and production, the devices will be made using photolithographic construction techniques. A genomic sequencing device capable of generating raw DNA sequence at less than $0.01 total costs per base is targeted for completion in five years. The proposal has four Program Goals: 1) Design, construct, and evaluate integrated DNA analysis systems. A system-level approach will be used to evaluate all components and fabrication technologies developed by the Program Project. Success will be measured at the level of complete genotyping and sequencing system. (Project 1) 2) Design, construct, and evaluate improved microfluidics, microreactions, and microseparations. Multistep handling and reactions will be tested on microfabricated devices. Advanced gel electrophoresis systems for sequencing t read-length >500 bp will be designed and tested. (Project 2) 3) Develop improved fabrication methods and optical sensors for integrated microsystems. Thin film deposited polymer and micromolded polymer will be examined for use as substrates and water-tight channels. Optical sensors will be designed and tested for improved sensitivity and stability. (Project 3) 4) Provide facilities and support for construction and testing of complex, integrated microfluidic devices. A central assembly and testing facility will be maintained that can rapidly build and test prototypes and incremental improvements for each of the projects (Core Unit).